Tools
Tools
Mingers, Toni, Barthels, Stefan, Mass, Violetta, Acuña, José Manuel Borrero de, Biedendieck, Rebekka, Cooke, Ana, Dailey, Tamara A., Gerdes, Svetlana, Blankenfeldt, Wulf, Dailey, Harry A., Warren, Martin J. 
ORCID: https://orcid.org/0000-0002-6028-6456, Jahn, Martina and Jahn, Dieter
(2024)
The alternative coproporphyrinogen III oxidase (CgoN) catalyzes the oxygen-independent conversion of coproporphyrinogen III into coproporphyrin III.
Frontiers in Microbiology, 15.
ISSN 1664-302X
Preview |
PDF (fmicb-15-1378989)
- Published Version
Available under License Creative Commons Attribution. Download (3MB) | Preview |
Abstract
Nature utilizes three distinct pathways to synthesize the essential enzyme cofactor heme. The coproporphyrin III-dependent pathway, predominantly present in Bacillaceae, employs an oxygen-dependent coproporphyrinogen III oxidase (CgoX) that converts coproporphyrinogen III into coproporphyrin III. In this study, we report the bioinformatic-based identification of a gene called ytpQ, encoding a putative oxygen-independent counterpart, which we propose to term CgoN, from Priestia (Bacillus) megaterium. The recombinantly produced, purified, and monomeric YtpQ (CgoN) protein is shown to catalyze the oxygen-independent conversion of coproporphyrinogen III into coproporphyrin III. Minimal non-enzymatic conversion of coproporphyrinogen III was observed under the anaerobic test conditions employed in this study. FAD was identified as a cofactor, and menadione served as an artificial acceptor for the six abstracted electrons, with a KM value of 3.95 μmol/L and a kcat of 0.63 per min for the substrate. The resulting coproporphyrin III, in turn, acts as an effective substrate for the subsequent enzyme of the pathway, the coproporphyrin III ferrochelatase (CpfC). Under aerobic conditions, oxygen directly serves as an electron acceptor, but is replaced by the more efficient action of menadione. An AlphaFold2 model of the enzyme suggests that YtpQ adopts a compact triangular shape consisting of three domains. The N-terminal domain appears to be flexible with respect to the rest of the structure, potentially creating a ligand binding site that opens and closes during the catalytic cycle. A catalytic mechanism similar to the oxygen-independent protoporphyrinogen IX oxidase PgoH1 (HemG), based on the flavin-dependent abstraction of six electrons from coproporphyrinogen III and their potential quinone-dependent transfer to a membrane-localized electron transport chain, is proposed.
| Item Type: | Article |
|---|---|
| Additional Information: | Data availability statement: The original contributions presented in the study are included in the article/Supplementary material, further inquiries can be directed to the corresponding author. Funding Information: The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was funded by various grants of the Deutsche Forschungsgemeinschaft (DFG). |
| Uncontrolled Keywords: | alternative heme biosynthesis,anaerobic metabolism,bacillaceae,coproporphyrinogen iii oxidase,priestia megaterium,microbiology,microbiology (medical) ,/dk/atira/pure/subjectarea/asjc/2400/2404 |
| Faculty \ School: | Faculty of Science |
| UEA Research Groups: | Faculty of Medicine and Health Sciences > Research Centres > Norwich Institute for Healthy Aging |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 23 Oct 2024 15:30 |
| Last Modified: | 16 Dec 2024 01:43 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/97158 |
| DOI: | 10.3389/fmicb.2024.1378989 |
Downloads
Downloads per month over past year
Actions (login required)
 |
View Item |